Methods for producing phenylalanine derivatives having a quinazolinedione skeleton and intermediates for production thereof

ABSTRACT

The present invention provides a method for producing a phenylalanine derivative(s) having a quinazolinedione ring of formula (5), including steps comprising of reacting an acylphenylalanine derivative(s) of formula (1) with a carbonyl group-introducing reagent(s) and a derivative(s) of anthranilic acid to form an asymmetric urea intermediate(s); making the asymmetric urea intermediate(s) into a quinazolinedione compound(s) of formula (4) in the presence of a base(s); and N-alkylating quinazolinedione ring amide of the obtained quinazolinedione compounds with N-alkylation agents. This production method is an industrially applicable method for producing phenylalanine derivatives having a quinazolinedione skeleton, which are compounds highly useful as drugs having α 4 integrin inhibiting activity. In the formulae (1) and (5), R1 represents a phenyl group having a substituent(s) and the like, R2 represents an alkyl group and the like, R3 represents a dialkylamino group and the like, and R4 represents an alkyl group and the like.

CROSS REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.11/207,751, filed Aug. 22, 2005, which, in turn, is a continuation ofInternational Application No. PCT/JP04/001982, filed Feb. 20, 2004, thedisclosures of which are incorporated herein by reference in theirentireties. This application claims priority to Japanese PatentApplication No. 2003-042560, filed Feb. 20, 2003, the disclosure ofwhich is incorporated by reference in its entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to methods for producing phenylalaninederivatives having a quinazolinedione skeleton, which are compoundshighly useful as drugs having α 4 integrin inhibiting activity, andintermediates thereof.

2. Discussion of the Background

Recently, research on inflammatory diseases in which α 4integrin-depending adhesion process participates in the pathology suchas rheumatoid arthritis, inflammatory bowel diseases, systemic lupuserythematosus, multiple sclerosis, Sjogren's syndrome, asthma,psoriasis, allergy, diabetes, cardiovascular diseases, arterialsclerosis, restenosis, tumor proliferation, tumor metastasis andtransplantation rejection has been advanced, and application to treatingor preventing agents of the compounds having α 4 integrin inhibitingactivity has been expected.

The applicant has already invented new phenylalanine derivatives havingα 4 integrin inhibiting activity, which are highly useful as treating orpreventing agents for inflammatory diseases in which α 4integrin-depending adhesion process participates in the pathology andfiled a patent application (WO2002-16329).

Conventionally, as the method for producing the phenylalaninederivatives having a quinazolinedione skeleton, it has been reportedsuch as that a quinazolinedione skeleton is constructed via amideintermediates by supporting phenylalanine derivatives on a solid-phaseresin (WO2002-16329 and Synlett, 3, 333-336, 2001).

However, though the solid-phase synthesis method has excellentadvantages as synthesis of a wide range of derivatives, generally it isnot a method suitable for industrialization because the amount ofderivatives which can be supported on a solid-phase resin haslimitations and, as a result, the amount of the object substance whichcan be prepared at one time is extremely small. Further, in thesolid-phase synthesis, a reaction reagent(s) is generally excessivelyused, and this is inappropriate from the point of industrialization.

In addition, by substituting the solid-phase synthesis method with aliquid-phase method based on the solid-phase synthesis method, forexample, in accordance with well-known examples of reports (S. M.Gadekar, et al., J. Am. Chem. Soc. 4666-4667, 1964, and L. Gouillex, etal., Tetrahedron lett., 37(39), 7031, 1996), a quinazolinedione skeletoncan be constructed by a synthesizing method comprising steps of reactingamine with carboxylic acid of anthranilic acid to form an amide, andreacting an amino group of anthranilic acid with ethyl chloroformate,1,1′-carbonyldiimidazole or the like to make it in carbamate orcarbonylimidazolyl form and then forming a quinazolinedione ring with abase(s). However, when the compound is synthesized, it has problems inthat the number of reaction processes are large and therefore the yieldis low.

On the other hand, as the method via urea intermediates, it has beenknown such as that amine is reacted with isocyanate to form an urea anda quinazolinedione ring is formed with a base (for example, WO2002-16329and B. Taub, J. Org. Chem., 26, 5238-5239, 1961).

However, isocyanate is typically a liquid with a pungent odor and highlytoxic, and it is known that isocyanate sometimes inducesself-polymerization to produce isocyanurate or reacts with water in theair to disintegrate. Thus, isocyanate is typically low in chemicalstability and has toxicity.

From these mentioned above, it is needed to find methods for producingphenylalanine derivatives having a quinazolinedione skeleton suitablefor industrialization.

SUMMARY OF THE INVENTION

The object of the present invention is to provide industriallyapplicable methods for producing phenylalanine derivatives having aquinazolinedione skeleton, which are compounds highly useful as drugproducts having α 4 integrin inhibiting activity.

The present invention also intends to provide intermediates forproduction of the phenylalanine derivatives having a quinazolinedioneskeleton.

The inventors studied the above problems to be solved and they foundindustrially applicable production methods in which phenylalaninederivatives having a quinazolinedione skeleton are conducted in highyield by a synthesizing method using a carbonyl group-introducingreagent(s) via urea intermediates, which is concise operation in mildreaction temperature without complicated solvent extraction andconcentration. The present invention has been completed on the basis ofthis finding.

[1] Namely, the present invention provides methods for producingphenylalanine derivatives having a quinazolinedione ring of followingformula (5), comprising steps of:

reacting an acylphenylalanine derivative(s) of formula (1):

wherein R1 represents a phenyl group which may have a substituent(s) ora pyridyl group which may have a substituent(s), R2 represents an alkylgroup which may have a substituent(s), and the derivative(s) is in asalt(s) with chemically acceptable acid(s) or free form(s),

with a carbonyl group-introducing reagent(s) and an anthranilic acidderivative(s) of formula (2):

wherein R3 represents a dialkylamino group, a monoalkylamino group, anamino group, a hydrogen atom, a halogen atom, an alkyl group,perfluoroalkyl group, an alkoxy group, a nitro group, an alkyl groupsubstituted with a dialkylamino group, an alkyl group substituted with amonoalkylamino group, an alkyl group substituted with an amino group, analkyl group substituted with an alkenyl group, an alkyl groupsubstituted with an alkynyl group, a carboxyl group, an alkoxycarbonylgroup, an alkylthio group or an arylthio group; R4 represents a hydrogenatom, an alkyl group or a benzyl group which may have a substituent(s);and R5 represents an alkyl group or an alkylcarbonyl group, and thederivative(s) is in a salt(s) with chemically acceptable acid(s) or freeform(s), to form an asymmetric urea intermediate(s) of formula (3):

wherein R1 to R5 are defined above;

making the asymmetric urea intermediate(s) into a quinazolinedionecompound(s) of formula (4) in the presence of a base(s):

wherein R1 to R4 are defined above; and

when R4 represents a hydrogen atom in the obtained quinazolinedionecompound(s) of the formula (4), N-alkylating a quinazolinedione ringamide of the quinazolinedione compound(s) with an N-alkylation agent(s)to form the phenylalanine derivative(s) having a quinazolinedioneskeleton of formula (5):

wherein R1 to R3 are defined above, R4 represents an alkyl group or abenzyl group which may have a substituent(s).

The present invention also provides the following compounds of (1) to(5), which are intermediates for production of the phenylalaninederivative(s) having a quinazolinedione ring of formula (5).

(1) Methylester of N^(α)-(2,6-dichlorobenzoyl)-4-amino-L-phenylalaninewherein, in the formula (1), R1 represents a 2,6-dichlorophenyl groupand R2 represents a methyl group, and salts thereof with chemicallyacceptable acids.

(2) Methylester of 5-dimethylamino-2-aminobenzoic acid wherein, in theformula (2), R3 represents a dimethylamino group, R4 represents ahydrogen atom and R5 represents a methyl group, and salts thereof withchemically acceptable acids.

(3) Methylester of2-(3-{4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureide)-5-dimethylaminobenzoic acid wherein, in the formula(3), R1 represents a 2,6-dichlorophenyl group, R2 represents a methylgroup, R3 represents a dimethylamino group, R4 represents a hydrogenatom and R5 represents a methyl group, and salts thereof with chemicallyacceptable acids.

(4) The compound wherein, in the formula (2), R3 represents adialkylamino group, a monoalkylamino group, an alkyl group substitutedwith a dialkylamino group, an alkyl group substituted with amonoalkylamino group, an alkyl group substituted with an alkynyl group,a carboxyl group, an alkoxycarbonyl group or an alkylthio group, R4represents a hydrogen atom and R5 represents a methyl group, and saltsthereof with chemically acceptable acids.

(5) The compound wherein, in the formula (3), R1 represents a2,6-dichlorophenyl group, R2 represents a methyl group, R3 represents adialkylamino group, a monoalkylamino group, an alkyl group substitutedwith a dialkylamino group, an alkyl group substituted with amonoalkylamino group, an alkyl group substituted with an alkynyl group,a carboxyl group, an alkoxycarbonyl group or an alkylthio group, R4represents a hydrogen atom and R5 represents a methyl group, and saltsthereof with chemically acceptable acids.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following embodiments are preferable in the present invention.

[2] The production method according to above [1], wherein, in theformulae (2) to (5), R3 represents a dialkylamino group, amonoalkylamino group, an alkyl group substituted with a dialkylaminogroup, an alkyl group substituted with a monoalkylamino group or analkyl group substituted with an alkynyl group.

[3] The production method according to above [1], wherein, in theformulae (2) to (5), R3 represents a dialkylamino group, amonoalkylamino group, an amino group, a hydrogen atom, a halogen atom,an alkyl group, a perfluoroalkyl group, an alkoxy group or a nitrogroup.

[4] The production method according to above [1], wherein, in theformulae (2) to (5), R3 represents a dialkylamino group.

[5] The production method according to any one of above [1] to [4],wherein the carbonyl group-introducing reagent(s) is1,1′-carbonyldiimidazole or chloroformate.

[6] The production method according to any one of above [1] to [5],wherein the base is potassium carbonate or sodium methoxide.

[7] The production method according to any one of above [1] to [6],wherein the N-alkylation agent is methyl p-toluenesulfonate.

[8] The production method according to above [1], comprising steps ofreacting a carbonyl group-introducing reagent selected from the groupconsisting of 1,1′-carbonyldiimidazole and chloroformate and thecompound of the formula (2) wherein R3 represents a dimethylamino group,R4 represents a hydrogen atom and R5 represents a methyl group with thecompound of the formula (1) wherein R1 represents a 2,6-dichlorophenylgroup and R2 represents a methyl group to obtain methylester of2-(3-{4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureid-e)-5-dimethylaminobenzoicacid; converting it in the presence of potassium carbonate or sodiummethoxide into methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-(6-dimethylamino-2,4[1H,3H]-quinazolinedione-3-yl)-L-phenylalanine of the formula (4); and thenN-alkylating the resultant with methyl p-toluenesulfonate to obtainmethylester ofN^(α)-(2,6-dichlorobenzoyl)-4-(1-methyl-6-dimethylamino-2,4[1H,3H]-quinazolinedione-3-yl)-L-phenylalanine.

[9] The production method according to above [1], comprising steps ofreacting a carbonyl group-introducing reagent selected from the groupconsisting of 1,1′-carbonyldiimidazole and chloroformate and thecompound of the formula (2) wherein R3 represents a dimethylamino group,R4 represents a methyl group and R5 represents a methyl group with thecompound of the formula (1) wherein R1 represents a 2,6-dichlorophenylgroup and R2 represents a methyl group to obtain the compound of theformula (3); converting it in the presence of potassium carbonate orsodium methoxide into methylester ofN^(α)-(2,6-dichloro-benzoyl)-4-(1-methyl-6-dimethylamino-2,4[1H,3H]-quinazolinedione-3-yl)-L-phenylalanine.

Next, the compounds in the present invention are described as follows.

R1 represents a phenyl group which may have a substituent(s) or apyridyl group which may have a substituent(s). In this connection,examples of the substituents are a halogen atom, an alkyl group, ahalogenoalkyl group including a perfluoroalkyl group, an alkoxy group, ahalogenoalkoxy group including a perfluoroalkoxy group, an alkylthiogroup, a nitro group, an alkylsulfonylamino group and a tetrazolylgroup. Here, an alkyl group as a component preferably has 1 to 6 carbonatoms and particularly preferably 1 to 3 carbon atoms, and they may besame or different from each other. R1 is preferably a phenyl groupsubstituted with a halogen atom and/or an alkyl group, and, for example,they are preferably 2,6-dichlorophenyl group, 2,6-dimethylphenyl group,2-chloro-6-methylphenyl group, 2-chlorophenyl group, 2-methylphenylgroup, 2,4,6-trichlorophenyl group, 2,4,6-trimethylphenyl group and2,6-dichloro-4-methylphenyl group.

R2 is an alkyl group which may have a substituent(s). In thisconnection, an alkyl group preferably has 1 to 6 carbon atoms andparticularly preferably 1 to 3 carbon atoms.

When R2 has a substituent(s), such substituent(s) include a substitutedor unsubstituted lower alkylcarbonyloxy group, a substituted orunsubstituted lower alkoxycarbonyloxy group, a substituted orunsubstituted amino group, a lower alkoxy group, a halogen atom, asubstituted or unsubstituted aryl group, a substituted or unsubstitutedheteroaryl group and a substituted or unsubstituted carbamoyl group.

Here, when the substituent(s) of R2 is a lower alkylcarbonyloxy group, alower alkoxycarbonyloxy group or a lower alkoxy group, alkyl and alkoxythereof preferably have 1 to 6 carbon atoms, and include chain, cyclic,linear and branched ones.

Further, when the substituent(s) of R2 is an aryl group, it represents amono- or bi-cyclic aromatic hydrocarbon group having 6 to 10 carbonatoms. For example, it includes a phenyl group and a naphthyl group.When the substituent(s) of R2 is a heteroaryl group, it represents a 5to 8 membered mono-, bi- or tricyclic aromatic heterocyclic groupincluding 1 to 4 hetero atoms selected from the group consisting of anoxygen atom, a sulfur atom and a nitrogen atom as a cyclic atom. Forexample, they include a pyridyl group, a pyridazinyl group, a pyrimidylgroup, a pyrazinyl group, a furyl group, a thienyl group, a pyrrolylgroup, an isoxazolyl group, an oxazolyl group, an isothiazolyl group, athiazolyl group, a pyrazolyl group, an imidazolyl group, a tetrazolylgroup, an indolyl group, a benzimidazolyl group, a quinolyl group and anisoquinolyl group. Here, a substituent(s) of the aryl group and thehetero aryl group is, for example, a halogen atom, an alkoxy group, analkyl group, a hydroxy group, a halogenoalkyl group and a halogenoalkoxygroup. Among these, a pyridyl group, a furyl group and a thienyl groupare preferred.

Meanwhile, when the substituent(s) of R2 is a lower alkylcarbonyloxygroup or a lower alkoxycarbonyloxy group, the substituent(s) thereofinclude a lower alkyl group, a lower alkenyl group, a lower alkoxygroup, a hydroxy group, an amino group and an amino group substitutedwith a lower alkyl group including monosubstitution or disubstitutionthereof. A methyl group and an ethyl group are preferred among these.

When the substituent(s) of R2 is an amino group, the substituent(s)thereof include a lower alkyl group, a lower alkoxycarbonyl group and alower alkylsulformyl group. Among these, a methyl group and an ethylgroup are preferable. Here, two substituents may bond together to form aring and, when forming a ring, they may also sandwich an oxygen,nitrogen or sulfur atom between them. For example, a substituted aminogroup includes a cyclic amino group such as 1-piperidinyl group and4-morphonyl group, a cyclic amide group such as 2-oxo-1-pyrrolidinylgroup and a cyclic urea group such as 2-oxoimidazoline-1-yl group and2-oxoimidazolidine-1-yl group.

Further, when the substituent(s) of R2 is an aryl group or a heteroarylgroup, the substituent(s) thereof include a halogen atom, an alkoxygroup, an alkyl group, a hydroxy group, a halogenoalkyl group and ahalogenoalkoxy group.

When the substituent(s) of R2 is a carbamoyl group, the substituent(s)thereof include a lower alkyl group and a phenyl group, and mono- anddi-substitutions thereof are also included.

When R2 has a substituent(s), the substituent(s) thereof is preferably alower alkylcarbonyloxy group, a chlorine atom, a pyridyl group, a furylgroup, a thienyl group and a lower dialkylcarbamoyl group.

In the formula (1), an amino group is preferably in the para positionamong oltho, meta and para positions on a benzene ring.

R3 represents a dialkylamino group, a monoalkylamino group, an aminogroup, a hydrogen atom, a halogen atom, an alkyl group, a perfluoroalkylgroup, an alkoxy group, a nitro group, an alkyl group substituted with adialkylamino group, an alkyl group substituted with a monoalkylaminogroup, an alkyl group substituted with an amino group, an alkyl groupsubstituted with an alkenyl group, an alkyl group substituted with analkynyl group, a carboxyl group, an alkoxycarbonyl group, an alkylthiogroup or an arylthio group, and R4 represents a hydrogen atom, an alkylgroup or a benzyl group which may have a substituent(s). While R5 may bewhatever OR5 is removed from COOR5 by a base, an alkyl group and analkylcarbonyl group are preferable. In R3 to R5, an alkyl group as acomponent preferably has 1 to 6 carbon atoms and particularly preferably1 to 3 carbon atoms.

R3 are preferably a dialkylamino group, a hydrogen atom, a halogen atom,a monoalkylamino group, an alkyl group substituted with a dialkylaminogroup, an alkyl group substituted with a monoalkylamino group, an alkylgroup substituted with an alkynyl group, a carboxyl group, analkoxycarbonyl group or an alkylthio group. Particularly preferred onesare a dialkylamino group, a monoalkylamino group, an alkyl groupsubstituted with a dialkylamino group, an alkyl group substituted with amonoalkylamino group, an alkyl group substituted with an alkynyl group,a carboxyl group, an alkoxycarbonyl group and an alkylthio group.

A dialkylamino group represents an amino group disubstituted with analkyl group having 1 to 6 carbon atoms, including a cyclic one.Preferably it is an amino group disubstituted with an alkyl group having1 to 3 carbon atoms or a cyclic amino group having 2 to 6 carbon atoms.For example, it includes a dimethylamino group, a diethylamino group, amethylethylamino group, a pyrrolidinyl group, a piperidyl group, adipropylamino group, a methylpropylamino group and an ethylpropylaminogroup.

A monoalkylamino group represents an amino group monosubstituted with analkyl group having 1 to 6 carbon atoms, including alkylamino group witha cyclic alkyl group(s). Preferably it is an amino group monosubstitutedwith an alkyl group having 1 to 4 carbon atoms such as a methylaminogroup, an ethylamino group, a propylamino group, an isopropylaminogroup, a butylamino group and a cyclopropylmethylamino group.

An alkyl group substituted with a dialkylamino group is an alkyl grouphaving 1 to 6 carbon atoms, which is substituted with the samesubstituent(s) as those of the dialkylamino group. Preferably it is analkyl group having 1 to 3 carbon atoms, which is substituted with thesame substituent(s) as those of the dialkylamino group. For example, itincludes either of a methyl group, an ethyl group or a propyl groupsubstituted with a dimethylamino group, a diethylamino group, amethylethylamino group, a pyrrolidinyl group, a piperidyl group, adipropylamino group, a methylpropylamino group or an ethylpropylaminogroup. Particularly preferable ones are a dimethylaminomethyl group, adiethylaminomethyl group, a methylethylaminomethyl group and the like.

An alkyl group substituted with a monoalkylamino group is an alkyl grouphaving 1 to 6 carbon atoms, which is substituted with the samesubstituent(s) as those of the monoalkylamino group. Preferably it is analkyl group having 1 to 3 carbon atoms, which is substituted with thesame substituent(s) as those of the monoalkylamino group. For example,it includes either of a methyl group, an ethyl group or a propyl groupsubstituted with a methylamino group, an ethylamino group, a propylaminogroup, an isopropylamino group, a butylamino group or acyclopropylmethylamino group. Particularly preferable ones are amethylaminomethyl group, an ethylaminomethyl group, a methylaminoethylgroup, an ethylaminoethyl group and the like.

An alkyl group substituted with an amino group is an alkyl group having1 to 6 carbon atoms, which is substituted with an amino group, andpreferably an alkyl group having 1 to 3 carbon atoms, which issubstituted with an amino group. For example, it includes an aminomethylgroup, an aminoethyl group, an aminopropyl group and the like.

An alkyl group substituted with an alkenyl group is an alkyl grouphaving 1 to 6 carbon atoms, which is substituted with an alkenyl grouphaving 2 to 6 carbon atoms, and preferably an alkyl group having 1 to 3carbon atoms, which is substituted with an alkenyl group having 2 to 4carbon atoms. For example, it includes —CH2CH═CH2, —CH2CH2CH═CH2 and thelike.

An alkyl group substituted with an alkynyl group is an alkyl grouphaving 1 to 6 carbon atoms, which is substituted with an alkynyl grouphaving 2 to 6 carbon atoms, and preferably an alkyl group having 1 to 3carbon atoms, which is substituted with an alkynyl group having 2 to 4carbon atoms. For example, it includes —CH2C═CH, —CH2CH2C═CH and thelike.

An alkoxycarbonyl group represents an alkoxycarbonyl group having 2 to 7carbon atoms and preferably having 2 to 4 carbon atoms, such as amethoxycarbonyl group, an ethoxycarbonyl group and a propyloxycarbonylgroup.

An alkylthio group represents a thio group substituted with an alkylgroup having 1 to 6 carbon atoms and preferably a thio group substitutedwith an alkyl group having 1 to 3 carbon atoms, such as a methylthiogroup, an ethylthio group and a propylthio group.

An arylthio group represents a phenylthio group and a naphthylthiogroup.

Particularly, R3 is preferably a dimethylamino group, a diethylaminogroup, a methylethylamino group, a pyrrolidinyl group, a piperidylgroup, a methylamino group, an ethylamino group, a propylamino group, acyclopropylmethylamino group, a dimethylaminomethyl group, adiethylaminomethyl group, a dimethylaminoethyl group, adiethylaminoethyl group, a methylaminomethyl group, an ethylaminomethylgroup, a propylaminomethyl group, a methylaminoethyl group, anethylaminoethyl group, a propylaminoethyl group, HC≡CCH2 group, acarboxyl group, a methoxycarbonyl group, an ethoxycarbonyl group, amethylthio group or an ethylthio group.

R4 is preferably a hydrogen atom or an alkyl group. In the formula (2),(3) and (4), a hydrogen atom is further preferable and, in the formula(5), an alkyl group is further preferable. While a substituent(s) of abenzyl group includes an alkyl group, an alkoxy group, a halogen atomand the like, an unsubstituted benzyl group is preferable.

R5 is particularly preferably an alkyl group.

In the formula (2), it is preferable that R3 is in the para position toan amino group.

The synthetic intermediates of the formula (1) are synthesized asfollows.

A phenylalanine having a nitro group on an aromatic ring and acidchloride are condensed under the conditions of the well-known method,Schotten-Baumann reaction (such as N. O. V. Sonntag, Chem. Rev. 52, 272,1953) to produce acylphenylalanine derivatives. Then, said carboxylicacid is esterified using the well known method (such as R. C. LarockComprehensive Organic Transformations (2nd Ed.), p. 1932-1941,Wiley-VCH, New York) to synthesize alkylester of acylphenylalanine. Theconventional method, catalytic reduction with a transition metalcatalyst(s) (such as R. C. Larock Comprehensive Organic Transformations(2nd Ed.), p. 821-828, Wiley-VCH, New York) is applied to thesynthesized substance in the presence of hydrogen gas to obtain thecorresponding compounds of the formula (1).

For example, as the method for producing methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine which is a newsynthetic intermediate wherein, in the formula (1), R1 represents a2,6-dichlorophenyl group and R2 represents a methyl group, said compoundcan be synthesized by following processes: condensing a publicly-knownand commercially available 4-nitro-L-phenylalanine and a similarlypublicly-known and commercially available 2,6-dichlorobenzoylchloride;subjecting the resultant to methyl esterification; and reduction of anitro group.

Concretely, 4-nitro-L-phenylalanine and 2,6-dichlorobenzoylchloride arecondensed in a mixed solvent of a sodium hydroxide aqueous solution andacetone with keeping the reaction temperature at 5 to 15° C., and thencrystallization is conducted to obtain a corresponding condensedsubstance, N^(α)-(2,6-dichlorobenzoyl)-4-nitro-L-phenylalanine almostquantitatively.

Subsequently, the condensed substance is suspended into methanol, andmethyl esterification is conducted by heating with adding concentratedsulfuric acid. Then, crystallization is conducted to obtain acorresponding methyl esterified substance, methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-nitro-L-phenylalanine almostquantitatively.

Then, catalytic reduction reaction is conducted to the methyl esterifiedsubstance using a transition metal catalyst(s) of a nitro group which isa well-known method (such as F. S. Dovell et al., J. Am. Chem. Soc., 87,2767, 1965) and preferably using a platinum carbon catalyst poisonedwith sulfur, and hydrogen gas. Thereafter, crystallization is conductedto obtain methylester ofNα-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine in high yield, wherein,in the formula (1), R1 represents a 2,6-dichlorophenyl group and R2represents a methyl group.

The yield in case of going through the three steps from4-nitro-L-phenylalanine is about 90%.

Meanwhile, the above-mentioned synthesizing method is not a sole methodfor producing the compound of the formula (1) and does not limit themethod of the present invention. Similarly, the yield indicates anaverage value thereof and doe not limit the value.

On the other hand, as the method for producing the compound of theformula (2), said compound can be obtained by well-known methods fromcommercially available trisubstituted benzene derivatives.

For example, as the method for producing methylester of5-dimethylamino-2-aminobenzoic acid which is a new syntheticintermediate wherein, in the formula (2), R3 represents a dimethylaminogroup, R4 represents a hydrogen atom and R5 represent a methyl group,said compound can be synthesized by following processes: dimethylamination, methyl esterification and reduction of a nitro group from apublicly-known and commercially available 5-chloro-2-nitrobenzoic acid.

Concretely, 5-chloro-2-nitrobenzoic acid is dissolved in a dimethylamineaqueous solution and heated to produce 5-dimethylamino-2-nitrobenzoicacid. Hydrochloric acid is added to the reaction solution to precipitateout the product as a solid material. The solid material is separated toobtain 5-dimethylamno-2-nitrobenzoic acid almost quantitatively.

Next, the 5-dimethylamno-2-nitrobenzoic acid is dissolved inconcentrated sulfuric acid/methanol and heated to conduct methylesterification in order to produce methylester of5-dimethylamino-2-nitrobenzoic acid, which is a well-known method.Toluene and water are added to the reaction solution, and the obtainedproduct is extracted to an organic layer, concentrated, and crystallizedto obtain 5-dimethylamino-2-nitrobenzoic acid in high yield.

Next, catalytic reduction is conducted to a nitro group of methylesterof 5-dimethylamino-2-nitrobenzoic acid under the acidic condition byhydrochloric acid, using transition metal catalysts such as palladiumcarbon and hydrogen gas in a methanol solvent. Crystallization isconducted to obtain methylester of 5-dimethylamino-2-aminobenzoicacid/dihydrochloride in high yield.

The yield via the three processes from 5-chloro-2-nitrobenzoic acid isabout 80%.

Further, thus obtained methylester of 5-dimethylamino-2-amino benzoicacid of the formula (2) can be obtained stably as salts with acidicsubstances, e.g., hydrochloride.

However, the above-mentioned synthesizing method is not a sole methodfor producing the compound of the formula (2) and does not limit theproduction method of the present invention. Similarly, the yieldindicates an average value thereof and does not limit the value.

The step 1 of the production method of the present invention isexplained below.

The method for producing phenylalanine derivatives having aquinazolinedione ring of the formula (5) in the present invention is asfollows. The compound of the formula (1) which is the importantintermediate and the compound of the formula (2) are converted intoasymmetric urea intermediates by using a carbonyl group-introducingreagent(s), preferably 1,1′-carbonyldiimidazole or chloroformate, andthen the compound of the formula (3) is converted to the compound of theformula (4) under mild basic condition.

In case of the compound wherein R4 in the formula (4) represents ahydrogen atom, the compound can be prepared without isolating thecompound of the formula (4), by successively conducting N-alkylationreaction with N-alkylation agents under basic condition. It may be alsopossible to isolate the compound of the formula (4) and then conductN-alkylation reaction.

For example, an amino group of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine of the formula (1)is converted into carbonylimidazolyl by 1,1′-carbonyldiimidazole usingan organic solvent(s) having suitable solubility such as, particularlypreferably, acetonitrile. Without isolating the intermediates thereof,methylester of 5-dimethylamino-2-aminobenzoic acid of the formula (2) isput into the reaction solvent to be able to obtain methylester of2-(3-{4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureide)-5-dimeth-ylaminobenzoicacid of the formula (3), which is asymmetric urea intermediates, in highyield.

In the present specification, “a carbonyl group-introducing reagent(s)”indicates those wherein only a carbonyl group in atomic groups of aquinazolinedione ring is derived from the present reagent. For instance,they include 1,1′-carbonyldiimidazole (Organic syntheses collectivevolume V, P. 201-204, Wiley, New York, 1973, as a synthetic examplethereof), chloroformate and the like. These are publicly known andcommercially available.

Further, it is also possible to use reagents wherein an imidazolyl groupof 1,1′-carbonyldiimidazole is substituted with other heteroarylremoving group(s), such as 1,1′-carbonyldi(1,2,4-triazole) (animidazolyl group in this reagent is substituted with a triazoyl groupand the reagent is commercially available). The substituents thereof arenot limited to only an imidazolyl group and a triazoyl group. It is alsopossible to use other heteroaryl removing groups as the substituents.

It is further possible to use N,N′-disuccinimidyl carbonate(DSC)(N-hydroxysuccinimide in this reagent is a removing group and thereagent is commercially available).

Chloroformate includes reagents having 2 to 10 carbon atoms, such asphenyl chloroformate, nitrophenyl chloroformate, methoxyphenylchloroformate, methyl chloroformate, ethyl chloroformate, isobutylchloroformate, octyl chloroformate and benzyl chloroformate, though theyare not limited to these examples.

Similarly, it is also possible to use phosgene and phosgene analoguessuch as triphosgene as the carbonyl group-introducing reagents. Theseare gas or liquid and highly toxic (Reference: RTECS SY 5600000) ascompared with the above reagents and therefore difficult to deal with.Further, since a particular kind of facilities is usually required anddistribution thereof is limited, these reagents are not very favorablein the carbonyl group-introducing reagents.

1,1′-carbonyldiimidazole is particularly preferred as a carbonylgroup-introducing reagents. When 1,1′-carbonyldiimidazole is used, it issuperior in that the amount of produced by-products is small and theobjective asymmetric urea intermediates are obtained in high yield.

A carbonyl group-introducing reagents is preferably used in 0.8 to 1.2mol equivalent weight to 1 mol of the compound of the formula (1).

The compound of the formula (2) is preferably used in 0.8 to 1.2 molequivalent weight to 1 mol of the compound of the formula (1).

The solvents of the present reaction include organic solvents havingsuitable solubility to the compounds of the formula (1) such asmethylester of N^(α)-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine. Forexample, it is possible to use acetonitrile, tetrahydrofuran (THF),N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), halogenatedcarbon hydride such as methylene chloride, pyridine, pyrrolidone,N-methylpyrrolidone, or mixed solvents thereof.

Above all, when acetonitrile is used, products can be easily separatedfrom the reaction solvent by filtration. Therefore, it is possible toisolate very pure asymmetric urea intermediates of the formula (3) byonly a simple separation with filtration. Since it does not require anycumbersome operations such as extraction and concentration of solvents,it is useful as industrialized process. Similarly, it is also possibleto obtain crystals of the objective compound by the steps of thereaction with N,N-dimethylformamide (DMF); adding poor solvents such asalcohols having 1 to 10 carbon atoms or water to precipitate asymmetricurea intermediates as a solid substance; and then separation byfiltration thereof. As a result, acetonitrile and N,N-dimethylformamideare particularly preferable as the solvents of the present reaction.

The concentration of the above reaction is preferably that applicable asindustrialized process. For example, when acetonitrile is used as areaction solvent, the reaction should be conducted in 1 to 0.01M andparticularly preferably around 0.2M, from the point of flowability instirring the reaction solution or crystallizing solution.

When the compound of the formula (1) is converted intocarbonylimidazolyl by carbonyl group-introducing reagents such as1,1′-carbonyldiimidazole, the reaction temperature thereof is preferablywithin the range of around 0° C. to not above the boiling point of thereaction solvent. To conduct the reaction around 0° C. to not above 10°C. is more industrially preferable in that it is useful for inhibitingside reactions and improving the yield. The reaction time is preferablearound 1 to 5 hours.

In the condensation reaction of the compound of the formula (1)converted into carbonylimidazolyl with methylester of5-dimethylamino-2-aminobenzoic acid of the formula (2), the reactiontemperature thereof is preferably within the range of around 0° C. tonot above the boiling point of the applied solvent. Particularly, thereaction is more preferably conducted at the reaction temperature ofaround 50° C. since urea bond formation reaction is completed in around2 to 3 hours and the asymmetric urea intermediates of the formula (3)can be obtained in high yield.

However, the reaction temperature and time are not limited to the aboveand the reaction time is determined by the balance with the reactiontemperature. It is desirable from the industrial point of view that thereaction solution is controlled by analytical methods such as HPLC.

In the above reaction, input order of raw materials and reagents is notparticularly limited. However, the method wherein the compound of theformula (1) is first reacted with a carbonyl group-introducingreagent(s) to convert into carbonylimidazolyl and then reacted with thecompound of the formula (2) is more preferable from the point of thehigh yield and side reactions as compared with the method wherein thecompound of the formula (2) is first converted into carbonylimidazolyl.While, in the production method of the present invention, the compoundof the formula (2) may be first converted into carbonylimidazolyl, andthe compound of the formula (1), a carbonyl group-introducing reagent(s)and the compound of the formula (2) may be reacted simultaneously.

Next, described herein is the step 2.

The asymmetric urea intermediates of the formula (3) form aquinazolinedione ring in the presence of a base in a suitable reactionsolvent to give the quinazolinedione compounds of the formula (4).

The “base” herein includes an inorganic base and organic base. Inorganicbases include salts with alkali metals such as potassium carbonate,sodium carbonate, cesium carbonate, sodium methoxide, sodium ethoxideand the like; and salts with alkaline earth metals such as calciumcarbonate and magnesium carbonate. Organic bases include triethylamine,ethanolamine, morpholine, piperidine, dicyclohexylamine,1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) andN,N-diisopropyl-N-ethylamine (DIPEA). Inorganic bases are preferable andparticularly potassium carbonate and sodium methoxide are preferable.

The base is preferably used in 0.1 to 2.0 mol equivalent weight to 1 molof the compound of the formula (3), and it is more preferable that 1 molequivalent weight or less of a catalyst is used. The reaction time ispreferably 1 to 6 hours.

For example, when potassium carbonate is used as a base, 0.1 to 2 molequivalent weight thereof is preferable and 0.2 to 0.4 mol equivalentweight is more preferable. When sodium methoxide is used as a base, 0.1to 1.0 mol equivalent weight thereof is preferable and 0.2 to 0.4 molequivalent weight is more preferable. 1 to 2 hours of the reaction timeare enough at that time.

The solvents in the above reactions may be those in which the compoundof the formula (3) is dissolved and the reaction proceeds smoothly. Forexample, they include dimethylformamide (DMF) and a mixed solvent withalcohols containing dimethylformamide as a main ingredient, though thesolvents are not limited to these.

Meanwhile, when potassium carbonate is used as a base, it is preferableto use a mixed solvent of DMF and methanol from the point of shorteningof the reaction time. As the relative proportions of the mixed solventof DMF and methanol, around 10 to 1 is suitable, though the proportionsare not limited to this.

As the reaction concentration of the above reactions, it is preferablethat the reaction is conducted in the concentration applicable asindustrialized process of within 0.01 to 2M and, for example, around0.25M in case of the mixed solvent of DMF and methanol, though theconcentration thereof is not limited to these.

As the reaction temperature, 0° C. to not above the boiling point of thesolvent, and preferably around 25° C. is suitable.

The quinazolinedione compounds of the formula (4) produced by the abovereactions can be precipitated as solid substance by adding water or anaqueous solution of hydrochloric acid dropwise to the reaction solution,or by adding the reaction solution to water or an aqueous solution ofhydrochloric acid, and then precipitated substance can be isolated bythe typical separating methods.

When potassium carbonate is used as a base, the quinazolinedione ringformation reaction is conducted to the asymmetric urea intermediates ofthe formula (3) and then N-methylation reaction can be soon linked afterthat without isolating the compound of the formula (4) and, therefore,one process in the reaction processes can be skipped. When takingaccount of improvement in production efficiency from the point ofindustrialization, this method is particularly useful as industrializedprocess since the isolating process of the compound of the formula (4)can be simplified.

Finally, described herein is the step 3.

In the formula (4), when R4 is a hydrogen atom, the compound can bederived into quinazolinedione compounds of the formula (5) withN-alkylation agents in the presence of a base.

Meanwhile, it is also possible to isolate the quinazolinedione compoundsof the formula (4), which is produced in the step 2, and thenN-alkylate. However, it is preferable from the point ofindustrialization to N-alkylate without isolation. In the presentspecification, “N-alkylation agents” indicate reagents which canintroduce an alkyl group on a nitrogen atom, and haloalkane, alkylsulfonate and benzyl halide which may be substituted are included, forexample.

Here, haloalkane and alkyl sulfonate are preferably those having 1 to 10carbon atoms. Those having 1 to 6 carbon atoms are further preferableand those having 1 to 3 carton atoms are particularly preferable.Haloalkane includes, for example, methyl iodide and ethyl iodide, andalkyl sulfonate includes, for example, methyl methanesulphonate, ethylmethanesulphonate, methyl ethanesulphonate, ethyl ethanesulphonate,methyl p-toluenesulphonate and ethyl p-toluenesulphonate. Benzyl halideincludes benzylchloride, benzylbromide and the like, and thesubstituents thereof are an alkyl group, an alkoxy group, a halogen atomand the like.

For example, in the production of the compound wherein R4 in the formula(5) is a methyl group, methyl p-toluenesulphonate is suitable from thepoint of industrialization. Namely, methyl p-toluenesulphonate has thehigher boiling point as compared with methyl iodide and is easy to dealwith under room temperature. Further, methyl p-toluenesulphonate has thefavorable flowability of the reaction solution and is suitable for theindustrialized process with solution sending.

As the usage amount of N-alkylation agents, the range of 1 to 10 molequivalent weight thereof and preferably around 1.2 to 2.0 molequivalent weight is suitable to the compound of the formula (3) or (4).The amount of the reagents can be increased or decreased in accordancewith progress of the reaction.

A base includes inorganic bases and organic bases. Here, examples ofinorganic bases are salts with alkali metals such as potassiumcarbonate, sodium carbonate, cesium carbonate, sodium methoxide, sodiumethoxide and the like; and salts with alkaline earth metals such ascalcium carbonate and magnesium carbonate. Organic bases includetriethylamine, ethanolamine, morpholine, piperidine, dicyclohexylamine,1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) andN,N-diisopropyl-N-ethylamine (DIPEA). Inorganic bases are preferable andparticularly potassium carbonate is preferable.

The base is preferably used in 1.5 to 2 mol equivalent weight to thecompound of the formula (3) or (4) and more preferably around 1.8 molequivalent weight, while the amount is not limited to these and can beincreased or decreased in accordance with progress of the reaction.

The reaction solvents may be those in which the compound of the formula(3) or (4) is dissolved and the reaction proceeds smoothly. For example,they include dimethylformamide (DMF) and a mixed solvent with alcoholscontaining dimethylformamide as a main ingredient, though the solventsare not limited to these. Meanwhile, when potassium carbonate is used asa base, it is preferable to use a mixed solvent of DMF and methanol fromthe point of shortening of the reaction time. As the relativeproportions of the mixed solvent of DMF and methanol, around 10 to 1 issuitable, though the proportions are not limited to this. As thereaction concentration of the above reactions, it is preferable that thereaction is conducted in the concentration applicable as industrializedprocess of within 0.01 to 2M and, for example, around 0.25M in case ofthe mixed solvent of DMF and methanol, though the concentration thereofis not limited to these.

As the reaction temperature, 0° C. to not above the boiling point of thereaction solvent, and preferably around 40° C. is suitable, though thetemperature thereof is not limited to these. The reaction time may be 3to 18 hours, and it is desirable from the industrial point of view thatthe reaction solution is administered by analytical methods such asHPLC.

For example, an asymmetric urea intermediate wherein, in the formula(4), R1 is 2,6-dihlorophenyl group, R2 is a methyl group and R3 is adimethylamino group is dissolved at 25° C. in a mixed solvent of DMF andmethanol. The quinazolinedione ring formation reaction is conducted bystirring for 2 hours using 0.2 mol equivalent weight of potassiumcarbonate to the compound of the formula (4). Then, 1.5 mol equivalentweight of methyl p-toluenesulphonate and 1.8 mol equivalent weight ofpotassium carbonate are added to conduct N-methylation reaction. Twoprocesses are gone through from the formula (4) to be able to obtainmethylester ofN^(α)-(2,6-dichlorobenzoyl)-4-(1-methyl-6-dimethylamino-2,4[1H,3H]-quinazolinedione-3-yl)-L-phenylalanine of the formula (5) in theyield of 80 to 90%.

The compound of the formula (5) can be precipitated as solid substanceby adding water to the reaction solvent and isolated by the typicalseparating methods such as filtration, though the method is not limitedto this.

The above production methods describe the case using the compoundwherein, in the formula (2), R4 is a hydrogen atom. As for the compoundswherein, in the formula (2), R4 is an alkyl group or a benzyl groupwhich may have a substituent(s), as the reaction process withN-alkylation agents is not necessary, the compound of the formula (1)and the compound of the formula (2) are derived into asymmetric ureaintermediates of the formula (3) by using a carbonyl group-introducingreagent(s) and preferably 1,1′-carbonyldiimidazole; the compound of theformula (3) is converted to the compound of the formula (4) under mildbasic condition; and then the compound of the formula (4) is isolated toobtain the objective compound of the formula (5) (because the formula(4) and the formula (5) are identical in this case).

From the above production methods, herein provided is the industriallyapplicable method for producing phenylalanine derivatives having aquinazolinedione ring of the formula (5) via the crucial intermediate(3) from crucial intermediates of the formulae (1) and (2).

Meanwhile, when R3 is a monoalkylamino group, an amino group, an alkylgroup substituted with a monoalkylamino group or an alkyl groupsubstituted with an amino group, a hydrogen atom(s) directly bonding toa nitrogen atom constituting the amino group may be protected in advanceby a proper protecting group(s) and then the protection thereof may beremoved by suitable methods of removal of protection. The methods forprotection and removal of protection are described in, for example,“Protecting group in organic synthesis” by Theodora W. Greene, Peter G.M. Wuts, Second edition, John Wiley & Sons Inc., New York, 1991,309-385.

For example, a hydrogen atom(s) directly bonding to a nitrogen atomconstituting the amino group may be substituted, by using acylationagents in ordinary acylation methods, with an alkylcarbonyl group having2 to 7 carbon atoms which may have a substituent(s) such as an acetylgroup, a propionyl group, a butyryl group, an isobutyryl group, avaleryl group, an isovaleryl group, a pivaloyl group, a trifluoroacetylgroup and the like; an arylcarbonyl group which may have asubstituent(s) such as a benzoyl group; an arylalkylcarbonyl group whichmay have a substituent(s) such as a benzylcarbonyl group; analkoxycarbonyl group having 2 to 7 carbon atoms which may have asubstituent(s) such as a methoxycarbonyl group, an ethoxycarbonyl group,a propyloxycarbonyl group, a butoxycarbonyl group, atertiary-butoxycarbonyl group, a trifluoromethoxy carbonyl group and thelike; an aryloxycarbonyl group which may have a substituent(s) such as aphenoxycarbonyl group; and an arylalkyloxycarbonyl group which may havea substituent(s) such as a benzyloxycarbonyl group.

In this case, after completion of the reaction of the first, second orthird step in the present invention, particularly after the completionof the N-alkylation reaction in the third step of the present invention,a protecting group(s) can be removed under the acidic condition or thebasic condition and the like in case of an alkylcarbonyl group having 2to 7 carbon atoms which may have a substituent(s), an arylcarbonyl groupwhich may have a substituent(s) and an arylalkylcarbonyl group which mayhave a substituent(s); under the basic condition and the like in case ofan alkoxycarbonyl group having 2 to 7 carbon atoms which may have asubstituent(s) and an aryloxycarbonyl group which may have asubstituent(s); under the acidic condition and the like in case of atertiary-butoxycarbonyl group; and under the catalytic reductioncondition (hydrogenating reaction) and the like.

Further, for instance, a hydrogen atom(s) directly bonding to a nitrogenatom constituting the amino group may be substituted, by usingbenzylation reagents in ordinary benzylation methods, with an arylalkylgroup which may have a substituent(s) such as a benzyl group, aphenethyl group, a methylbenzyl group, a methoxybenzyl group and ahalobenzyl group, and the like. In this case, after completion of thereaction of the first, second or third step in the present invention,particularly after the completion of the N-alkylation reaction in thethird step of the present invention, a protecting group(s) can beremoved under the catalytic reduction condition (hydrogenating reaction)and the like.

Meanwhile, the substituents in case of the above protecting groups“which may have a substituent(s)” include, for example, a halogen atom,an alkoxy group, an alkyl group, a hydroxy group, a halogenoalkyl groupand a halogenoalkoxy group.

The “halogen atom” in the present specification indicates a fluorineatom, a chlorine atom, a bromine atom and an iodine atom. The“halogeno-” as the component in a substituent indicates fluoro-,chloro-, bromo- and iodo-.

The “chemically acceptable acids” in the formula (1) or (2) includeinorganic acids such as hydrochloric acid, sulfuric acid, phosphoricacid, nitric acid and hydrobromic acid; organic carboxylic acid such asacetic acid, citric acid, benzoic acid, maleic acid, fumaric acid,tartaric acid, succinic acid, trifluoroacetic acid, tannic acid, butyricacid, hibenzic acid, pamoic acid, enanthic acid, decanoic acid, teoclicacid, salicylic acid, lactic acid, oxalic acid, mandelic acid and malicacid; and organic sulfonic acid such as methanesulfonic acid,p-toluenesulfonic acid and benzenesulfonic acid. The free forms areparticularly preferable in the compound of the formula (1) and thehydrochloride is particularly preferable in the compound of the formula(2). They may be hydrates or solvates thereof.

In the present invention, when R3 in the formula (5) is a dialkylaminogroup, the resulting compound is therapeutically superior, andtherefore, R3 in the formulae (2) and (3) are preferably dialkylaminogroups. Among dialkylamino groups, a dimethylamino group, a diethylaminogroup, a methylethylamino group, a pyrrolidinyl group and a piperidinylgroup are preferable. Particularly, it is preferable that R3 is adimethylamino group.

Similarly, it is also preferable that R3 in the formulae (2) and (3) area monoalkylamino group, an alkyl group substituted with a dialkylaminogroup, an alkyl group substituted with a monoalkylamino group, an alkylgroup substituted with an alkynyl group, a carboxyl group, analkoxycarbonyl group or an alkylthio group. Particularly preferred onesare a methylamino group, an ethylamino group, a propylamino group, acyclopropylmethylamino group, a dimethylaminomethyl group, adiethylaminomethyl group, a dimethylaminoethyl group, adiethylaminoethyl group, a methylaminomethyl group, an ethylaminomethylgroup, a propylaminomethyl group, methylaminoethyl group,ethylaminoethyl group, a propylaminoethyl group, HC≡CCH2 group, acarboxyl group, a methoxycarbonyl group, an ethoxycarbonyl group, amethylthio group and an ethylthio group.

Among these, when R3 in the formulae (2) and (3) are a monoalkylaminogroup or an alkyl group substituted with a monoalkylamino group, ahydrogen atom(s) directly bonding to a nitrogen atom constituting theamino group may be substituted with an alkylcarbonyl group having 2 to 7carbon atoms which may have a substituent(s) such as an acetyl group, apropionyl group, a butyryl group, an isobutyryl group, a valeryl group,an isovaleryl group, a pivaloyl group, a trifluoroacetyl group and thelike; an arylcarbonyl group which may have a substituent(s) such as abenzoyl group; an arylalkylcarbonyl group which may have asubstituent(s) such as a benzylcarbonyl group; an alkoxycarbonyl grouphaving 2 to 7 carbon atoms which may have a substituent(s) such as amethoxycarbonyl group, an ethoxycarbonyl group, a propyloxycarbonylgroup, a butoxycarbonyl group, a tertiary-butoxycarbonyl group, atrifluoromethoxy carbonyl group and the like; an aryloxycarbonyl groupwhich may have a substituent(s) such as a phenoxycarbonyl group; and anarylalkyloxycarbonyl group which may have a substituent(s) such as abenzyloxycarbonyl group. A hydrogen atom(s) directly bonding to anitrogen atom constituting the amino group may be substituted with anarylalkyl group which may have a substituent(s) such as a benzyl group,a phenethyl group, a methylbenzyl group, a methoxybenzyl group and ahalobenzyl group, and the like. The substituents in case of the aboveprotecting groups “which may have a substituent(s)” include a halogenatom, an alkoxy group, an alkyl group, a hydroxy group, a halogenoalkylgroup and a halogenoalkoxy group.

EXAMPLES

Next, Examples will further illustrate the present invention in detail.The following Examples only explain the present invention and do notparticularly limit the invention.

<Analysis Conditions>

TMS was used as an internal reference material in ¹H and ¹³C NMR, andmeasurement was conducted with AVANCE 400 mHz NMR of Bruker BioSpinGmbH. As DMSO-d₆, the product (containing 0.03% TMS) of Eurisotop CEAGroup was used. As an HPLC apparatus, LC10 series (Pump: LC-10AT,Controller: SCL-10A and Detector: SPD-10Avp) of Shimadzu Corporation wasused. As an autosampler, KMT-100× of Kyowaseimitsu Corporation (was used(the injection volume is 10 μl as long as it is not particularlydescribed). As an column oven, U-620 of Sugai Chemical Industry Co., LTDwas used. As a chromato waveform processing, C-R7A of ShimadzuCorporation was used. As raw materials and reagents in the presentExamples, commercial items thereof are used themselves withoutpurification.

<HPLC Analysis Conditions> Compositions of Solution A 0.1% TFA aqueoussolution eluting solvents: Solution B acetonitrile containing 0.1% TFAFlow volume: 1.0 mL/min. Detector: UV, 254 nm Used column reverse-phaseODS silica gel column (ODS-2 of GL Sciences, Inc.) Column size: innerdiameter of φ 4.6 mm and length of 150 mm Column temperature: 40° C.Gradient analysis (Solution A/Solution B) = beginning (90/10) − 25condition: minutes later (10/90) − 30 minutes later (10/90) Sampleinjection 10 μl volume:

Synthetic Reference Example of Synthetic Intermediate (1) Syntheticexample of the compound (6) Synthesis of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine

200 mL of water and 91 mL of acetone were mixed and 72.4 g (344 mmol) of4-nitro-L-phenylalanine was added thereto and cooled down to 10° C. orlower. 68 mL of 6M sodium hydroxide aqueous solution was added dropwiseto the solution so that the temperature thereof did not exceed 15° C.Keeping around pH 14, 73.58 g (344 mmol) of 2,6-dichlorobenzoyl chloridewas slowly added dropwise thereto. In order to keep around pH 14, ifneeded, a sodium hydroxide aqueous solution was added dropwise. 2 hourslater of completion of drop, 86 mL of 6M hydrochloric acid was addedwith keeping the temperature of the reaction solution at 15° C. or lowerto precipitate white crystals. After maturing at 10° C. or lower, thecrystals were separated, dried under reduced pressure at 60° C. toobtain 128.5 g of N^(α)-(2,6-dichlorobenzoyl)-4-nitro-L-phenylalanine.(Yield: 97%) ¹H NMR (400 MHz, DMSO-d₆): 9.12 (d, 1H, J=8.42 Hz), 8.16(d, 2H, J=8.78 Hz), 7.59 (d, J=8.77 Hz), 7.42 (m, 3H), 3.29 (m), 3.07(dd, 1H, J=3.64 and 10.42 Hz). ¹³C NMR (100 MHz, DMSO-d⁶): 172.25,163.74, 146.68, 146.18, 131.51, 131.37, 131.05, 128.37, 123.55, 53.17,36.74.

MS (FAB): m/z 383.1 (M+H)⁺

HRMS (FAB): m/z 383.0219 (M+H)⁺

Next, 117.3 g (306 mmol) ofN^(α)-(2,6-dichlorobenzoyl)-4-nitro-L-phenylalanine was added to 592 mLof methanol and dissolved. 31.6 g of 95% concentrated sulfuric acid wasadded dropwise being careful of heating. After the drop, the reactionwas conducted for 3 hours at the reaction temperature of 40° C. Afterconfirming completion of the reaction with HPLC, the reaction solutionwas cooled down to 30° C. or lower. 395 mL of water cooled down toaround 10° C. in advance was added dropwise in 1 hour so that thetemperature of the reaction solution did not exceed 30° C. Aftercrystallizing out, the reaction solution was matured for 5 hours withkeeping a crystallizing solution at 10° C. or lower. The crystals wereseparated by filtration, and then dried under reduced pressure at 60° C.to obtain 117.8 g of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-nitro-L-phenylalanine. (yield 97%)

¹H NMR (400 MHz, DMSO-d₆): 9.23 (d, 1H, J=8.2 Hz), 8.18 (d, 2H, J=8.8Hz), 7.59 (d, 2H, J=8.8 Hz), 7.38-7.46 (m, 3H), 4.88 (ddd, 1H, J=6.4,8.2 and 11 Hz), 3.69 (s, 3H), 3.31 (dd, 1H, J=6.4 and 14 Hz), 3.10 (dd,1H, J=11 and 14 Hz).

¹³C NMR (100 MHz, DMSO-d₆): 171.27, 163.83, 146.74, 145.80, 136.15,131.49, 131.05, 128.42, 123.58, 53.07, 52.40, 36.39.

MS (FAB): m/z 397.2 (M+H)⁺

HRMS (FAB): m/z 397.0345 (M+H)⁺

Further, 115.94 g (290 mmol) of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-nitro-L-phenylalanine and 28.37 g (0.5 mol% to substrate) of 3% platinum carbon powder (wet) were suspended to 825mL of methanol. Nitro groups in the suspended solution were reducedunder hydrogen gas atmosphere at 30° C. for 5 hours. After confirmingcompletion of the reaction with HPLC, the platinum catalyst was filteredout and the concentration of the reaction solution was adjusted. 539 mLof water was added dropwise thereto at the liquid temperature of around30° C., and crystallization by cooling was conducted at 10° C. or lower.The crystals were filtered out and dried under reduced pressure at 60°C. to obtain 84.61 g of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine. (yield 80%)

¹H NMR (400 MHz, DMSO-d₆): 9.23 (d, 1H, J=7.8 Hz), 7.38-7.47 (m, 3H),6.90 (d, 2H, J=7.0 Hz), 6.47 (d, 2H, J=7.0 Hz), 4.57 (ddd, 1H, J=5.8,7.8 and 9.1 Hz), 3.62 (s, 3H), 2.90 (dd, 1H, J=5.8 and 14 Hz), 2.79 (dd,1H, J=9.1 and 14 Hz).

¹³C NMR (100 MHz, DMSO-d₆): 171.87, 163.88, 147.52, 136.43, 131.65,131.34, 129.90, 128.34, 124.01, 114.14, 54.57, 52.07, 36.40.

MS (FAB): m/z 367.2 (M+H)⁺

HRMS (FAB): m/z 367.0585 (M+H)⁺

Synthetic Reference Example of Synthetic Intermediate (2) Syntheticexample of the compound of formula (7) Synthesis of methylester of2-amino-5-(dimethylamino) benzoic acid/dihydrochloride

30.0 g (148 mmol) of 5-chloro-2-nitrobenzoic acid was dissolved bystirring in 78 mL (744 mmol) of 50% dimethylamine aqueous solution undercooling in the ice bath. After the solution was put into apressure-resistant container and sealed, the solution was stirred byheating in the oil bath for 23 hours at 60° C. The reaction solution wassufficiently cooled down and the inner pressure thereof was released.After confirming completion of the reaction by HPLC analysis, thereaction solution was put into another container (using 50 mL of water),49.6 mL of concentrated hydrochloric acid, and then 200 mL of water wereadded thereto. Yellow crystals were precipitated by addition ofhydrochloric acid. The crystallizing solution was matured at 10° C.overnight, separated by filtration and dried under reduced pressure toobtain 30.95 g of 5-dimethylamino-2-nitrobenzoic acid. (yield 99%)

¹H NMR (400 MHz, DMSO-d₆): 8.88 (bs, 1H), 7.97 (d, 1H, J=9.4 Hz, arylcoupling=1.76 Hz), 6.78 (d, 1H, J=9.4 Hz, aryl coupling=2.84 and 1.92Hz), 6.71 (s, 1H, aryl coupling=2.88 and 1.60 Hz), 3.08 (s, 6H).

¹³C NMR (100 MHz, DMSO-d₆): 168.58, 153.86, 133.94, 132.85, 127.03,111.44, 109.69, 40.24.

MS (ESI): m/z 211.17 (M+H)⁺, 209.27 (M−H)⁻

Next, 40.0 g (190.30 mmol) of 5-dimethylamino-2-nitrobenzoic acid wassuspended in 160 mL of methanol at 25° C. This suspension was undercooled down in the ice bath and 53.6 mL of concentrated sulfuric acidwas added thereto. After adding the concentrated sulfuric acid, thetemperature of the solution rose up to about 30° C. The solution wasdirectly soaked in the bath of 60° C. and stirred by heating for 20hours. The progress of the reaction was confirmed by HPLC, and afterconfirming disappearance of a raw material, 400 mL of toluene was addedthereto to dilute. Then, 200 mL of water and an aqueous solution ofsodium hydroxide (wherein 38.06 g of sodium hydroxide was dissolved in200 mL of water) were added thereto. The aqueous layer was extractedwith 200 mL of toluene and a toluene solvent(s) was mixed together. Thetoluene layer was washed with 300 mL of saturated sodium bicarbonatewater. The toluene layer was further concentrated under reduced pressure(bath temperature: 50° C.) and adjusted so that the objective substancebecomes about 20 wt %. After distilling under reduced pressure, crystalsof the objective substance precipitated. After maturing them at roomtemperature for about one hour, 220 mL of n-heptane was added andfurther stirred at 5° C. overnight. The crystals were separated bysuction filtration and washed with 100 mL of n-heptane. These wetcrystals were dried under reduced pressure for 3 hours at 65° C. toobtain 34.82 g of methylseter of 5-dimethylamino-2-nitrobenzoic acid asyellow crystallized powder. (yield 82%)

¹H NMR (400 MHz, DMSO-d₆): 8.02 (d, 1H, J=9.4 Hz), 6.82 (d, 1H, J=9.36Hz, aryl coupling=2.56 Hz), 6.78 (s, 1H, aryl coupling=2.4 Hz), 3.83 (s,3H), 3.10 (s, 6H).

¹³C NMR (100 MHz, DMSO-d₆): 167.70, 153.92, 132.71, 132.34, 127.24,111.87, 110.07, 53.21, 40.28.

MS (FAB): m/z 224.24 (M)⁺

HR MS (FAB): m/z 224.0830 (M)⁺

Further, 10.06 g (44.9 mmol) of methylester of5-dimethylamino-2-nitrobenzoic acid was added to 50 mL of methanol andsuspended. 9.0 mL of 10M hydrochloric acid and 1.96 g (wet, 1 mol % tosubstrate) of 5% palladium carbon were added thereto. The reactioncontainer was substituted with hydrogen gas and stirred at roomtemperature overnight. A palladium catalyst was filtered out with Celiteand the filtrate was concentrated under reduced pressure by about a halfamount thereof. Then, 80 mL of acetone was added to the solution andconcentrated under reduced pressure three times to precipitate thecompound of the formula (12). The compound was further matured at 10° C.or lower and dried under reduced pressure to 11.16 g of methylester of2-amino-5-(dimethylamino) benzoic acid/dihydrochloride. (yield 93%)

¹H NMR (400 MHz, DMSO-d₆): 8.09 (s, 1H), 7.72 (d, 1H, J=9.0 Hz), 6.96(d, 1H, 9.08 Hz), 5.50 (bs), 3.83 (s, 3H), 3.04 (s, 6H).

¹³C NMR (100 MHz, DMSO-d₆): 167.12, 131.64, 126.66, 123.29, 118.7,108.88, 52.18, 45.84.

MS (FAB): m/z 195.3 (M+H)⁺

HR MS (FAB): m/z 195.1122 (M+H)⁺

Example 1

<Process>1

Synthetic example of the compound (8) Synthesis of methylester of2-(3-{-4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureid-e)-5-dimethylaminobenzoicacid

Method 1: In Case of Using 1,1′-carbonyldiimidazole (CDI) as a CarbonylGroup-Introducing Reagent

9.73 g (59.41 mmol) of 1,1′-carbonyldiimidazole was added to 310 mL ofacetonitrile and dissolved. The solution was cooled down to 10° C. orlower and 20.78 g (56.58 mmol) of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine of the formula (6)was added thereto and stirred. 2 hours later, 15.06 g (54.50 mmol) ofmethylester of 2-amino-5-(dimethylamino) benzoic acid/dihydrochloride ofthe formula (7) was added, heated up to 50° C. and stirred for 2 hours.After completion of the reaction, 62 mL of methanol was added to thereaction solution, and the solution was cooled down to 10° C. or lower.After maturing 10 hours or more, the crystals were separated byfiltration and dried under reduced pressure to obtain 30.03 g ofobjective methylester of2-(3-{-4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureid-e)-5-dimethylaminobenzoicacid of formula (8). (yield 88%)

¹H NMR (400 MHz, DMSO-d6): δ 9.56 (s, 1H), 9.46 (s, 1H), 9.22 (d, 1H,J=8.0 Hz), 8.07 (d, 1H, J=9.24 Hz), 7.47-7.38 (m, 5H), 7.19 (m, 3H),7.06 (m, 1H), 4.69 (m, 1H), 3.88 (s, 3H), 3.66 (s, 3H), 3.06 (dd, 1H,J=14.1 and 5.3 Hz), 2.93-2.88 (m, 1H), 2.88 (s, 6H)

¹³C NMR (100 MHz, DMSO-d6): δ 171.72, 168.20, 163.88, 152.87, 145.63,138.89, 136.38, 131.80, 131.64, 131.37, 130.46, 129.78, 128.36, 122.83,119.31, 118.54, 117.18, 112.99, 54.09, 52.59, 52.18, 40.80, 36.34.

MS (FAB): m/z 586.3 (M)⁺

HR MS (FAB): 586.1407 (M)⁺

Method 2: In Case of Using Phenyl Chloroformate as a CarbonylGroup-Introducing Reagent

2.08 g (5.45 mmol) of methylester ofN-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine of the formula (6) wasadded to 30 mL of acetonitrile and stirred at room temperature anddissolved. After under cooling in the ice bath, 0.83 mL of triethylamineand 0.72 mL (5.72 mmol) of phenyl chloroformate were added thereto.After warming the reaction solution to room temperature and stirring for1.5 hours, 1.46 g (5.45 mmol) of methylester of 2-amino-5-dimethylaminobenzoic acid/dihydrochloride and 1.51 mL of triethylamine were addedthereto and stirred at room temperature for 3 days. Precipitatedcrystals were filtered out, washed with methanol and dried under reducedpressure to obtain 2.55 g of a crystalline solid substance containingobjective methylester of2-(3-{4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureide)-5-dimeth-ylaminobenzoicacid of formula (8). (content 61.3 wt %, yield 49%)

Meanwhile, analytical data of the compound conformed to those of theabove mentioned Method 1.

Method 3: In Case of Using N,N′-disuccinimidyl carbonate (DSC) as aCarbonyl Group-Introducing Reagent

1.32 g (3.60 mmol) of methylester ofN-(2,6-dichlorobenzoyl)-4-amino-L-phenylalanine of the formula (6) wasadded to 15 mL of acetonitrile and stirred at room temperature anddissolved. 1.0 g (3.90 mmol) of N,N′-disuccinimidyl carbonate (DSC) wasadded to the solution and stirred at room temperature. 0.98 g (3.68mmol) of methylester of 2-amino-5-dimethylamino benzoicacid/dihydrochloride and 1.92 g of N,N-diisopropyl-N-ethylamine (DIPEA)were added to the reaction solution and stirred at 50° C. for 2.5 hours.The objective substance precipitated as a solid material as the reactionproceeded, and the suspension was cooled down to 10° C. or lower. Thecrystals were filtered out, washed with methanol and dried under reducedpressure to obtain 1.16 g of objective methylester of2-(3-{4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureide)-5-dimeth-ylaminobenzoicacid of formula (8). (yield 55%) Meanwhile, analytical data of thecompound conformed to those of the above mentioned Method 1.

<Process>2

Synthetic example of the compound of formula (9) Synthesis ofmethylester ofN^(α)-(2,6-dichlorobenzoyl)-4-(6-dimethylamino-2,4[1H,3H]-quinazo-linedione-3-yl)-L-phenylalanine

40.0 g (68.14 mmol) of methylester of2-(3-{-4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureid-e)-5-dimethylaminobenzoicacid of the formula (8) was added to 200 mL of N,N-dimethylformamide andstirred and dissolved at 25° C. 5.4 mL of 28% sodium methoxide/methanolsolution was added thereto and stirred at 25° C. for 2 hours. Aftercompletion of the reaction, the reaction solution was added dropwise to210 mL of an aqueous solution of hydrochloric acid to precipitate thecompound of formula (14). The compound was separated and dried underreduced pressure to obtain 36.74 g of the title compound. (yield 97.2%)

¹H NMR (400 MHz, DMSO-d₆): δ 11.20 (bs, 1H), 9.29 (d, 1H, J=8.12 Hz),7.47-7.38 (m, 5H), 7.29-7.26 (m, 1H), 7.18 (d, 2H, J=8.3 Hz), 7.12 (m,2H), 4.81 (m, 1H), 3.69 (s, 3H), 3.22 (dd, 1H, J=14.1 and 4.8 Hz), 3.02(dd, 1H, J=14.0 and 3.8 Hz), 2.91 (s, 6H).

¹³C NMR (100 MHz, DMSO-d₆): δ 171.70, 163.99, 162.75, 150.18, 146.80,137.15, 136.34, 134.80, 131.78, 131.36, 131.15, 129.84, 129.18, 128.32,122.05, 116.48, 115.03, 108.50, 53.70, 52.29, 40.93, 36.36.

MS (FAB): m/z 555.2 (M+H)⁺

HR MS (FAB): m/z 555.1172 (M+H)⁺

<Process>3

Synthetic Example of the Compound of Formula (10)

(Method 1) Synthesis of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-(1-methyl-6-dimethylamino-2,4[1H,3H-]-quinazolinedione-3-yl)-L-phenylalanineby N-methylation of the formula (9) [A production method in case ofisolating the compound of the formula (9)]

30.0 g (54.0 mmol) of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-(6-dimethylamino-2,4[1H,3H]-quinazo-linedione-3-yl)-L-phenylalanineof the formula (9) was added to a solution containing 180 mL ofN,N-dimethylformamide (DMF) and 20 mL of methanol and stirred anddissolved at 25° C. A DMF solution (20 mL as DMF) containing 15.3 g(81.1 mmol) of methyl of p-toluenesulfonic acid and 15.0 g (108.1 mmol)of potassium carbonate were added thereto. After addition thereof, thereaction solution was stirred at the reaction temperature of 40° C. for6 hours. Then, the solution was added dropwise being careful ofexothermic heating to an aqueous solution of hydrochloric acid (1.8 mLof 6M hydrochloric acid and 250 mL of water) which was cooled down inadvance at 10° C. or lower. The precipitated substance was filtered outand dried at 60° C. under reduced pressure to obtain 25.3 g of the titlecompound of the formula (10). (yield 82%)

(Method 2) Synthesis of methylester ofN^(α)-(2,6-dichlorobenzoyl)-4-(1-methyl-6-dimethylamino-2,4[1H,3H-]-quinazolinedione-3-yl)-L-phenylalanine[A production method in case of not isolating the compound of theformula (9)]

20 g (34.07 mmol) of methylester of2-(3-{4-[2-(2,6-dichlorobenzoylamino)-2-methoxycarbonylethyl]phenyl}ureid-e)-5-dimethylaminobenzoicacid of the formula (8) was stirred and dissolved in 110 mL ofN,N-dimethylformamide (DMF) at 20° C. 11 mL of methanol and 0.94 g (6.81mmol) of potassium carbonate were added thereto and stirred at 25° C.for 1 hour. After confirming completion of the quinazolinedione ringformation reaction with HPLC, without isolating the compound of theformula (9), a DMF solution (14 mL of DMF) containing 7.74 mL (51.11mmol) of methyl of p-toluenesulfonic acid and 8.46 g (61.33 mmol) ofpotassium carbonate were added to the reaction solution. N-methylationreaction was conducted to the solution at 40° C. After completion of thereaction, the reaction solution was added to water to precipitate thetitle compound of the formula (10) as a solid substance. Theprecipitated substance was filtered out and dried under reduced pressureto obtain 16.71 g thereof. (yield 86.2%)

¹H NMR (400 MHz, DMSO-d₆): δ 9.29 (d, 1H, J=8.12 Hz), 7.47-7.36 (m, 6H),7.32-7.29 (m, 1H), 7.24 (d, 1H, J=2.84 Hz), 7.18 (d, 2H, J=8.28 Hz),4.82 (m, 1H), 3.69 (s, 3H), 3.49 (s, 3H), 3.23 (dd, 1H, J=14.1 and 4.6Hz), 3.02 (dd, 1H, J=13.9 and 3.5 Hz), 2.94 (s, 6H).

¹³C NMR (100 MHz, DMSO-d₆): δ 171.73, 163.99, 161.88, 150.37, 146.73,137.20, 136.34, 135.34, 132.06, 131.78, 131.36, 129.89, 128.99, 128.32,121.34, 116.21, 116.00, 109.15, 53.65, 52.29, 40.75, 36.35, 30.88.

MS (ESI): m/z 569.33 (M+H)⁺

Anal. Calcd for C₂₈H₂₆N₄O₅Cl₂: C, 59.06; H, 4.60; N, 9.84; Cl, 12.45.Found: C, 59.08; H, 4.64; N, 9.82; Cl, 12.43.

Where a numerical limit or range is stated herein, the endpoints areincluded. Also, all values and subranges within a numerical limit orrange are specifically included as if explicitly written out.

Obviously, numerous modifications and variations of the presentinvention are possible in light of the above teachings. It is thereforeto be understood that, within the scope of the appended claims, theinvention may be practiced otherwise than as specifically describedherein.

All patents and other references mentioned above are incorporated infull herein by this reference, the same as if set forth at length.

1. Methylester of 5-dimethylamino-2-aminobenzoic acid according to the following formula (7)

or a salt thereof with a chemically acceptable acid.
 2. A hydrochloride salt of methylester of 5-dimethylamino-2-aminobenzoic acid. 